title: Development of a serum-free defined system employing growth factors for preantral follicle culture.
abstract: This study was conducted to evaluate if mouse preantral follicles can yield developmentally competent oocytes following culture in serum-free, defined medium. Donor follicles were obtained from 14-day-old B6CBAF1 mice, and cultured in α-MEM-Glutamax medium. The replacement of fetal bovine serum with knockout serum replacement (KSR) did not significantly reduce follicle growth or oocyte maturation in vitro, although it significantly reduced the development of oocytes after activation. Regardless of the replacement medium, follicle growth was not influenced by the addition of leukemia inhibitory factor (LIF). The addition of 100 ng/ml stem cell factor (SCF) to the KSR-supplemented serum-free medium significantly stimulated follicle development, which further improved blastocyst formation after oocyte activation. On Day 3 of culture, a significant increase in Bmp7 expression was detected in the SCF-containing medium compared with the serum-containing medium, whereas Gdf9 and Amh were increased in the serum-containing medium. A significant increase in estradiol production was detected under serum-free conditions, but minimal progesterone secretion was detected throughout the culture period. In conclusion, serum-free media can be used to optimize ovarian follicle cultures, and the addition of SCF is beneficial for deriving developmentally competent oocytes through follicle culture.
authors.affiliation: Stem Cell and Bioevaluation, WCU Biomodulation Program, Seoul National University, Seoul, Korea.
authors.forename: Young Hyun
authors.initials: YH
authors.lastname: Park
authors.forename: Seung Pyo
authors.initials: SP
authors.lastname: Gong
authors.forename: Hwa Young
authors.initials: HY
authors.lastname: Kim
authors.forename: Gil Ah
authors.initials: GA
authors.lastname: Kim
authors.forename: Jun Hee
authors.initials: JH
authors.lastname: Choi
authors.forename: Ji Yeon
authors.initials: JY
authors.lastname: Ahn
authors.forename: Jeong Mook
authors.initials: JM
authors.lastname: Lim
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D055419:Bone Morphogenetic Protein 7 / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D016895:Culture Media, Serum-Free / Q000737:chemistry*; D004958:Estradiol / Q000378:metabolism; D005260:Female; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects / Q000502:physiology*; D053605:Leukemia Inhibitory Factor / Q000494:pharmacology; D051379:Mice; D006080:Ovarian Follicle / Q000166:cytology / Q000187:drug effects / Q000254:growth & development*; D011374:Progesterone / Q000378:metabolism; D027724:Reproductive Techniques, Assisted; D019089:Stem Cell Factor / Q000494:pharmacology*
chemical_list: D055419:Bone Morphogenetic Protein 7; D016895:Culture Media, Serum-Free; D053605:Leukemia Inhibitory Factor; D019089:Stem Cell Factor; C525820:bmp7 protein, mouse; D011374:Progesterone; D004958:Estradiol